Nocardioides pakistanensis sp nov., isolated from a hot water spring of Tatta Pani in Pakistan

A Gram-staining positive, non-spore forming, non-pigmented and non-motile bacterium, designated as NCCP-1340(T), was isolated from a hot water spring, Tatta Pani, Pakistan. Cells of strain NCCP-1340(T) were observed to be aerobic, rod shaped, catalase and urease positive but H2S production and oxidase negative. Growth was observed at pH 6.0-8.0 (optimum pH 7.0) and at 20-40 A degrees C (optimum 37 A degrees C). The strain could tolerate 0-8 % NaCl (optimum 2 %, w/v). Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain NCCP-1340(T) belongs to the genus Nocardioides and is closely related to Nocardioides iriomotensis JCM 17985(T) (96.8 %), Nocardioides daedukensis KCTC 19601(T) (96.6 %), Nocardioides jensenii KCTC 9134(T) (96.1 %) and Nocardioides daejeonensis KCTC 19772(T) (96.1 %). The DNA-DNA relatedness values of strain NCCP-1340(T) with N. iriomotensis JCM 17985(T), N. daedukensis KCTC 19601(T) and N. jensenii KCTC 9134(T) were found to be less than 53 %. The DNA G+C content of strain NCCP-1340(T) was determined to be 71.8 mol %. The affiliation of strain NCCP-1340(T) to the genus Nocardioides was further supported by chemotaxonomic data which showed the presence of MK-8(H-4) as major menaquinone system; iso-C-16:0, C-17:0, C-16:0 10-methyl, iso-C-15:0 and C (15:0) as major cellular fatty acids; and diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and unidentified glycolipids and polar lipids in the polar lipids profile. The cell wall peptidoglycan contained LL-diaminopimelic acid as the diagnostic amino acid. On the basis of physiological and biochemical characteristics and the phylogenetic analyses, strain NCCP-1340(T) can be distinguished from the closely related taxa and thus represents a novel species of the genus Nocardioides, for which the name Nocardioides pakistanensis sp. nov. is proposed with the type strain NCCP-1340(T) (= DSM 29942(T) = JCM 30630(T)).